TFDG对IL-1β体外诱导大鼠软骨细胞炎性损伤的保护作用研究

周盈, 黄倩, 张甜, 陈萍

茶叶科学 ›› 2017, Vol. 37 ›› Issue (3) : 290-298.

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茶叶科学 ›› 2017, Vol. 37 ›› Issue (3) : 290-298.

TFDG对IL-1β体外诱导大鼠软骨细胞炎性损伤的保护作用研究

  • 周盈1, 黄倩2, 张甜1, 陈萍1,*
作者信息 +

Research on the Protective Effects of TFDG on IL-1β-induced Inflammatory Injury in Rat Chondrocytes in Vitro

  • ZHOU Ying1, HUANG Qian2, ZHANG Tian1, CHEN Ping1,*
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文章历史 +

摘要

体外分离培养SD雄性大鼠膝关节软骨细胞,经甲苯胺蓝及II型胶原(Col II)免疫荧光染色鉴定后,加入白介素-1β(IL-1β)诱导建立骨关节炎(OA)细胞模型,探讨茶黄素双没食子酸酯(TFDG)对OA软骨细胞的保护作用。细胞形态观察发现TFDG能明显改善OA软骨细胞形态。实时荧光定量PCR(Real-time PCR)结果显示,TFDG不仅可以上调软骨细胞分子标志物Col II mRNA的表达,还可以下调炎症因子IL-1β、IL-6 mRNA的表达。酶联免疫吸附实验(ELISA)检测结果进一步表明TFDG可明显降低炎症因子的分泌。免疫印迹(Western blot)检测结果证明,TFDG预干扰可降低炎症诱导酶环氧化酶COX-2蛋白表达量。这些结果说明TFDG通过减弱炎症反应,从而对IL-1β体外诱导大鼠软骨细胞炎性损伤起到保护作用。

Abstract

The chondrocytes were isolated and harvested from knee articular surface of male Sprague-Dawley rats, which were identified by toluidine blue and type II collagen immunofluorescence staining. To investigate potential protective effects of theaflavin-3,3′-digallate (TFDG) on IL-1β-induced inflammatory injury in rat chondrocytes in vitro, a model of osteoarthritis was established through stimulating rat chondrocytes with IL-1β. Inverted phase contrast microscopy showed TFDG treatment significantly improved osteoarthritis chondrocytes morphology. Real-time PCR results showed that TFDG treatment not only up-regulated chondrocyte marker Col II mRNA expression, but also down-regulated pro-inflammatory cytokines IL-1β and IL-6 mRNA expression. ELISA analysis further confirmed TFDG treatment significantly decreased the secretion of inflammatory factors. Western blot results showed that TFDG treatment significantly inhibited the inflammation-related enzyme COX-2 expression. Taken together, these results indicate that TFDG has a preventive effect on IL-1β-induced inflammatory injury in rat chondrocytes in vitro by reducing inflammatory reaction.

关键词

茶黄素双没食子酸酯 / 骨关节炎 / 软骨细胞 / 炎症

Key words

chondrocytes / inflammation / osteoarthritis / TFDG

引用本文

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周盈, 黄倩, 张甜, 陈萍. TFDG对IL-1β体外诱导大鼠软骨细胞炎性损伤的保护作用研究[J]. 茶叶科学. 2017, 37(3): 290-298
ZHOU Ying, HUANG Qian, ZHANG Tian, CHEN Ping. Research on the Protective Effects of TFDG on IL-1β-induced Inflammatory Injury in Rat Chondrocytes in Vitro[J]. Journal of Tea Science. 2017, 37(3): 290-298
中图分类号: Q946.84+1    R681.3   

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基金

国家茶叶产业体系(CARS-23)、国家自然科学基金(31200521)、高等学校博士学科点专项科研基金(20120101120112)

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